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Lipopolysaccharide/adenosine triphosphate-mediated signal transduction in the regulation of NLRP3 protein expression and caspase-1-mediated interleukin-1β secretion

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Abstract

Objective

Reactive oxygen species (ROS) plays a critical role in the regulation of NLRP3 inflammasome activation. However, the ROS-mediated signaling pathways controlling NLRP3 inflammasome activation are not well defined.

Methods

Using lipopolysaccharide (LPS) and adenosine triphosphate (ATP) activated murine macrophages as the testing model, cytokine release and protein expression were quantified by enzyme-linked immunosorbent assay and Western blot, respectively. ROS was scavenged by N-acetyl cysteine; NADPH oxidase, the major source of ROS, was inhibited by diphenyliodonium, apocynin or gp91-phox siRNA transfection; and protein kinase was inhibited by its specific inhibitor.

Results

LPS-induced NLRP3 protein expression was regulated through the NADPH oxidase/ROS/NF-κB-dependent, JAK2/PI3-kinase/AKT/NF-κB-dependent, and MAPK-dependent pathways, while ATP-induced caspase-1 activation was regulated through the NADPH oxidase/ROS-dependent pathway.

Conclusions

These results demonstrate that ROS regulates not only the priming stage, but also the activation stage, of NLRP3 inflammasome activation in LPS + ATP-activated macrophages.

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Acknowledgments

This work was supported by the National Science Council, Taiwan: contract grant numbers: NSC 98-2320-B-039-003-MY2 (KF Hua); 100-2313-B-197-002 (KF Hua).

Author information

Author notes

  1. Pei-Chun Liao

    Present address: Department of Cosmeceutics, China Medical University, Taichung, Taiwan

Authors and Affiliations

  1. Department of Biotechnology and Animal Science, National Ilan University, 1, Sec. 1, Shen-Lung Road, Ilan, 260, Taiwan

    Pei-Chun Liao, Ju-Ching Chou, Wei-Chih Dong, Chien-Nan Lin, Chai-Yi Lin & Kuo-Feng Hua

  2. Department of Cosmeceutics, China Medical University, Taichung, Taiwan

    Louis Kuoping Chao

  3. Division of Wood Cellulose, Taiwan Forestry Research Institute, Taipei, Taiwan

    Chen-Lung Ho

  4. Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan

    Ann Chen

  5. Graduate Institute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, Taiwan

    Shuk-Man Ka

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  1. Pei-Chun Liao
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Correspondence to Kuo-Feng Hua.

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Liao, PC., Chao, L.K., Chou, JC. et al. Lipopolysaccharide/adenosine triphosphate-mediated signal transduction in the regulation of NLRP3 protein expression and caspase-1-mediated interleukin-1β secretion. Inflamm. Res. 62, 89–96 (2013). https://doi.org/10.1007/s00011-012-0555-2

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  • DOI: https://doi.org/10.1007/s00011-012-0555-2

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